Search

Porous starch has been created through hydrolysis by amyloglucosidase and α-amylase. However, little information is known about the precise evolution of multi-scale structures of starch during digestion. In this study, rice starch and potato starch, containing different crystalline structures, were hydrolyzed by amyloglucosidase and α-amylase for 20 and 60 min, respectively, and their resulting structural changes were examined. The digestion process caused significant degradation of the molecular structures of rice and potato starches. In addition, the alterations in the ordered structures varied between the two starches. Rice starch exhibited porous structures, thicker crystalline lamellae as determined by small-angle X-ray scattering, and enhanced thermostability after digestion using differential scanning calorimetry. For rice starch, the extent of crystalline structures was analyzed with an X-ray diffractometer; it was found to first increase after 20 min of digestion and then decrease after 60 min of digestion. In contrast, potato starch did not display porous structures but exhibited thicker crystalline lamellae and a reduction in ordered structures after digestion. These findings suggest that it is possible to intentionally modulate the multi-scale structures of starch by controlling the digestion time, thereby providing valuable insights for the manipulation of starch functionalities. [ABSTRACT FROM AUTHOR]

This paper introduces an enzymatic approach to estimate internal mass-transfer resistances during food digestion studies. Cellulase has been used to degrade starch cell walls (where cellulose is a significant component) and reduce the internal mass-transfer resistance, so that the starch granules are released and hydrolysed by amylase, increasing the starch hydrolysis rates, as a technique for measuring the internal mass-transfer resistance of cell walls. The estimated internal mass-transfer resistances for granular starch hydrolysis in a beaker and stirrer system for simulating the food digestion range from 2.2 × 107 m−1 s at a stirrer speed of 100 rpm to 6.6 × 107 m−1 s at 200 rpm. The reaction rate constants for cellulase-treated starch are about three to eight times as great as those for starch powder. The beaker and stirrer system provides an in vitro model to quantitatively understand external mass-transfer resistance and compare mass-transfer and reaction rate kinetics in starch hydrolysis during food digestion. Particle size analysis indicates that starch cell wall degradation reduces starch granule adhesion (compared with soaked starch samples), though the primary particle sizes are similar, and increases the interfacial surface area, reducing internal mass-transfer resistance and overall mass-transfer resistance. Dimensional analysis (such as the Damköhler numbers, Da, 0.3–0.5) from this in vitro system shows that mass-transfer rates are greater than reaction rates. At the same time, SEM (scanning electron microscopy) images of starch particles indicate significant morphology changes due to the cell wall degradation. [ABSTRACT FROM AUTHOR]

Owing to the importance of α‐amylase in industry, it is necessary to develop a more simple and practical approach to measure α‐amylase activity. Herein, a fluorescence method was established for the detection of α‐amylase activity based on hydroxyproply‐β‐cyclodextrin‐curcumin inclusion complex (HP‐β‐CDs‐Cur). Curcumin was utilized as a signal probe, while hydroxyproply‐β‐cyclodextrin (HP‐β‐CDs) was employed both as a substrate of α‐amylase and the carrier of curcumin. α‐amylase is capable to hydrolyse α‐1,4‐glycosidic bond on the HP‐β‐CDs ring, facilitating the detection of its activity through the measurement of the decrease of fluorescence intensity caused by the release of curcumin into the aqueous. The results showed that there was a good linear relationship between fluorescence intensity and α‐amylase concentration in the range of 0.065~1.365 U mL−1, and the detection limit was 0.0065 U mL−1. The successful application of this method to evaluate the activities of multiple commercial α‐amylases underscores its robustness and reliability. [ABSTRACT FROM AUTHOR]

Summary: The effects of water content on the multi‐scale structure and digestibility of sweet potato starch (SPS) during heat‐moisture treatment (HMT) were studied, and the relationship between water content, starch structure and starch digestibility was analysed. The results showed that the surface of SPS granules changed after HMT, mainly in the form of depression, rupture and adhesion, and the change was more obvious with the increase of water content, and the crystalline structure changed from the original A‐type to A + V‐type. The relative crystallinity and double helix content decreased, the thickness of semi‐crystalline layered structure increased, and the peak intensity decreased. The molecular weight of SPS after HMT tended to be distributed in the low molecular weight range of 4 × 104 g mol−1 < Mw <4 × 105 g mol−1. These structural changes promoted the rearrangement of the intermolecular structure of SPS, which made amylase shield the original action site when acting on SPS, and converted part of rapidly digestible starch (RDS) content into slow digestible starch (SDS) and resistant starch (RS) content, resulting in the increase of SDS and RS content. Especially in HMT‐25, the RS content of SPS was the highest, which was 28.60%. The experimental data can provide theoretical support for broadening the application of SPS. [ABSTRACT FROM AUTHOR]

Thermophilic actinomycetes are commonly found in extreme environments and can thrive and adapt to extreme conditions. These organisms exhibit substantial variation and garnered significant interest due to their remarkable enzymatic activities. This study evaluated the potential of Streptomyces griseorubens NBR14 and Nocardiopsis synnemataformans NBRM9 strains to produce thermo-stable amylase via submerged fermentation using wheat and bean straw. The Box-Behnken design was utilized to determine the optimum parameters for amylase biosynthesis. Subsequently, amylase underwent partial purification and characterization. Furthermore, the obtained hydrolysate was applied for ethanol fermentation using Saccharomyces cerevisiae. The optimal parameters for obtaining the highest amylase activity by NBR14 (7.72 U/mL) and NBRM9 (26.54 U/mL) strains were found to be 40 and 30 °C, pH values of 7, incubation time of 7 days, and substrate concentration (3 and 2 g/100 mL), respectively. The NBR14 and NBRM9 amylase were partially purified, resulting in specific activities of 251.15 and 144.84 U/mg, as well as purification factors of 3.91 and 2.69-fold, respectively. After partial purification, the amylase extracted from NBR14 and NBRM9 showed the highest activity level at pH values of 9 and 7 and temperatures of 50 and 60 °C, respectively. The findings also indicated that the maximum velocity (Vmax) for NBR14 and NBRM9 amylase were 57.80 and 59.88 U/mL, respectively, with Km constants of 1.39 and 1.479 mM. After 48 h, bioethanol was produced at concentrations of 5.95 mg/mL and 9.29 mg/mL from hydrolyzed wheat and bean straw, respectively, through fermentation with S. cerevisiae. Thermophilic actinomycetes and their α-amylase yield demonstrated promising potential for sustainable bio-ethanol production from agro-byproducts. [ABSTRACT FROM AUTHOR]

In this study, 2(3),9(10),16(17),23(24)-tetrakis-[(N-methyl-(1-benzylpiperidin-4-yl)oxy)phthalocyaninato]zinc(II) iodide (ZnPc-2) was synthesized and characterized using spectral methods (FT-IR, 1H-NMR, UV-Vis and mass spectroscopy). The interaction of ZnPc-2 with DNA was investigated by using the UV/Vis titrimetric method, thermal denaturation profile, agarose gel electrophoresis and molecular docking studies. Additionally, the antidiabetic activity of ZnPc-2 was revealed spectroscopically by studying α-amylase and α-glucosidase inhibition activities. The spectroscopic results indicated that ZnPc-2 effectively binds to calf thymus-DNA (CT-DNA) with a Kb value of 7.5 × 104 M−1 and interacts with CT-DNA via noncovalent binding mode. Gel electrophoresis results also show that ZnPc-2 binds strongly to DNA molecules and exhibits effective nuclease activity even at low concentrations. Furthermore, docking studies suggest that ZnPc-2 exhibits a stronger binding tendency with DNA than the control compounds ethidium bromide and cisplatin. Consequently, due to its strong DNA binding and nuclease activity, ZnPc-2 may be suitable for antimicrobial and anticancer applications after further toxicological tests. Additionally, antidiabetic studies showed that ZnPc-2 had both α-amylase and α-glucosidase inhibition activity. Moreover, the α-glucosidase inhibitory effect of ZnPc-2 was approximately 3500 times higher than that of the standard inhibitor, acarbose. Considering these results, it can be said that ZnPc-2 is a moderate α-amylase and a highly effective α-glucosidase inhibitor. This suggests that ZnPc-2 may have the potential to be used as a therapeutic agent for the treatment of type 2 diabetes. [ABSTRACT FROM AUTHOR]

In this article, we focus on the plasma seed interaction and more specifically-on the feedback exerted by the seeds on the plasma properties. Dormant Arabidopsis seeds with different water contents (WC), namely 3%DW, 10%DW and 30%DW were exposed to cold ambient air plasma (C2AP) generated in a dielectric barrier device (DBD). It is found that increasing WC enhances the capacitive current of the DBD, generates a greater number of low energy streamers (characterized by current peaks lower than 10 mA) that preferentially interplay with the seeds. Since the resistive and capacitive components of the seeds modify the C2AP electrical properties, impedance measurements (also called LCRmetry) have been carried out to measure their main dielectric parameters before/after plasma exposure (seeds resistance, capacitance, complex relative permittivity, tangent loss and conductivity). It appears that WC significantly changes dielectric losses at low frequencies (<1 kHz) due to polarization relaxation of the polar molecules (i.e. water). LCRmetry further reveals that C2AP does not substantially alter seeds dielectric parameters, i.e. it neither adds or removes significant amounts of new materials, meaning that the relative starch, protein and lipid contents remain essentially unaffected. However, it cannot be discounted that some bulk properties of the Arabidopsis seeds may be modified, especially regarding their porosity. This characteristic could facilitate penetration of plasma-generated reactive oxygen species into the internal seed tissues, leading to the grafting of oxygenated groups. To corroborate this theory, water sorption isotherms have been achieved on Arabidopsis seeds and fitted with four thermodynamic models, including the Brunauer–Emmett–Teller model and the Generalized D'Arcy and Watt model. It is demonstrated that C2AP primarily strengthens water-seed affinity by modifying molecular interactions rather than changing the seed's moisture layer. This occurs despite a potential decrease in the number of adsorption sites, indicating a significant increase in overall seed hydrophilicity after plasma treatment. [ABSTRACT FROM AUTHOR]

Bacillus sp. PM06, previously isolated from sugarcane waste pressmud, could produce dual enzymes α-amylase and cellulase. The isolate's crude enzymes were purified homogeneously using ammonium sulfate precipitation followed by High Quaternary amine anion exchange chromatography. Purified enzymes revealed the molecular weights of α-amylase and cellulase as 55 and 52 kDa, with a purification fold of 15.4 and 11.5, respectively. The specific activity of purified α-amylase and cellulase were 740.7 and 555.6 U/mg, respectively. It demonstrated a wide range of activity from pH 5.0 to 8.5, with an optimum pH of 5.5 and 6.4 for α-amylase and cellulase. The optimum temperature was 50 °C for α-amylase and 60 °C for cellulase. The kinetic parameters of purified α-amylase were 741.5 ± 3.75 µmol/min/mg, 1.154 ± 0.1 mM, and 589 ± 3.5/(smM), using starch as a substrate. Whereas cellulase showed 556.3 ± 1.3 µmol/min/mg, 1.78 ± 0.1 mM, and 270.9 ± 3.8/(smM) of Vmax,Km, Kcat/Km, respectively, using carboxymethyl cellulose (CMC) as substrate. Among the various substrates tested, α-amylase had a higher specificity for amylose and CMC for cellulase. Different inhibitors and activators were also examined. Ca2+ Mg2+, Co2+, and Mn2+ boosted α-amylase and cellulase activities. Cu2+ and Ni2+ both inhibited the enzyme activities. Enzymatic saccharification of wheat bran yielded 253.61 ± 1.7 and 147.5 ± 1.0 mg/g of reducing sugar within 12 and 24 h of incubation when treated with purified α-amylase and cellulase. A more significant amount of 397.7 ± 1.9 mg/g reducing sugars was released from wheat bran due to the synergetic effect of two enzymes. According to scanning electron micrograph analysis, wheat bran was effectively broken down by both enzymes. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is a leading environmental issue worldwide. The current study was conducted to investigate Cd tolerance of 10 commercial white clover (Trifolium repens) cultivars during seed germination and to further explore differences in lipid remodelling, glycometabolism, and the conversion of lipids into sugars contributing to Cd tolerance in the early phase of seedling establishment as well as the accumulation of Cd in seedlings and mature plants. The results show that Cd stress significantly reduced seed germination of 10 cultivars. Compared to Cd‐sensitive Sulky, Cd‐tolerant Pixie accelerated amylolysis to produce more glucose, fructose, and sucrose by maintaining higher amylase and sucrase activities under Cd stress. Pixie maintained higher contents of various lipids, higher DGDG/MGDG ratio, and lower unsaturation levels of lipids, which could be beneficial to membrane stability and integrity as well as signal transduction in cells after being subjected to Cd stress. In addition, Pixie upregulated expression levels of key genes (TrACX1, TrACX4, TrSDP6, and TrPCK1) involved in the conversion of lipids into sugars for early seedling establishment under Cd stress. These findings indicate that lipid remodelling, enhanced glycometabolism, and accelerated conversion of lipids into sugars are important adaptive strategies for white clover seed germination and subsequent seedling establishment under Cd stress. In addition, Pixie not only accumulated more Cd in seedlings and mature plants than Sulky but also had significantly better growth and phytoremediation efficiency under Cd stress. Pixie could be used as a suitable and critical germplasm for the rehabilitation and re‐establishment of Cd‐contaminated areas. [ABSTRACT FROM AUTHOR]

AbstractAs compared with exogenous components, non-starch components (NSCS), such as proteins, lipids, non-starch polysaccharides (NSPs), and polyphenols, inherently present in cereals, are more effective at inhibiting starch digestibility. Existing research has mostly focused on complex systems but overlooked the analysis of the in-situ role of the NSCS. This study reviews the crucial mechanisms by which endogenous NSCS inhibit starch digestion, emphasizing the spatial distribution-function relationship. Starch granules are filled with pores/channels-associated proteins and lipids, embedding in the protein matrix, and maintained by endosperm cell walls. The potential starch digestion inhibition of endogenous NSCS is achieved by altering starch gelatinization, molecular structure, digestive enzyme activity, and accessibility. Starch gelatinization is constrained by endogenous NSCS, particularly cell wall NSPs and matrix proteins. The stability of the starch crystal structure is enhanced by the proteins and lipids distributed in the starch granule pores and channels. Endogenous polyphenols greatly inhibit digestive enzymes and participate in the cross-linking of NSPs in the cell wall space, which together constitute a physical barrier that hinders amylase diffusion. Additionally, the spatial entanglement of NSCS and starch under heat and non-heat processing conditions reduces starch accessibility. This review provides novel evidence for the health benefits of whole cereals. [ABSTRACT FROM AUTHOR]

Summary: In frozen part‐baked bread technology, starch is influenced by various processing factors. This study aimed to investigate the effect of part‐baking time, freezing rate and frozen storage time on pasting, thermal and structural properties of part‐baked bread. Wheat dough was part‐baked for 0, 3 or 6 min and frozen at −20 °C using two freezing rates: slow (0.15 °C min−1) or fast (1.45 °C min−1). Next, samples were stored in frozen conditions for 56 days and analysed every 14 days for starch damage, pasting properties, thermal and X‐ray analysis. The level of damaged starch increased with 3 min (69.3%), and 6 min (88.8%) of part‐baking time for both freezing rates compared to no part‐baked. In pasting analyses, peak viscosity decreased 27.3% and 38.4% for 3 and 6 min of part‐baking time, respectively, at 0 days of frozen storage; however, this tendency was observed for all frozen storage time. The gelatinisation enthalpy (ΔHg) decreased as part‐baking time increased, showing the maximum reduction at 6 min of part‐baked (94.9%). In X‐ray diffraction analyses at 3 and 6 min of part‐baking, the intensity of the signal increased 4.4% and 10.9%, respectively, probably due to the starch gelatinisation. These results can serve as a valuable guide for developing strategies to enhance the quality of part‐baked bread and improve production performance. [ABSTRACT FROM AUTHOR]

The vibrating superfine mill (VSM) is a machine that belongs to the micronization technique. In this study, VSM was employed to produce micronized tapioca starch by varying micronization times (15, 30, 45, and 60 min). The structural and physicochemical properties of the micronized starch were then examined. Scanning electron microscopy studies revealed that micronized starch was partially gelatinized, and the granule size dramatically increased when micronization time increased. X‐ray diffraction patterns showed that the relative crystallinity was decreased from 24.67% (native) to 4.13% after micronization treatment for 15 min and slightly decreased after that. The solubility of micronized starch significantly increased as the micronization time increased, which was associated with the destruction of the starch crystalline structure. Differential scanning calorimetry investigations confirmed that micronized starch was "partly gelatinized," and the degree of gelatinization increased to 81.27% when the micronization time was 60 min. The weight‐average molar mass was reduced by 15.0% (15 min), 30.9% (30 min), 55.7% (45 min), and 70.5% (60 min), respectively, indicating that the molecular structure was seriously degraded. The results demonstrated that the physicochemical changes of micronized starch granules were related to the destruction of the starch structure. These observations would provide details on micronized starch and its potential applications. Practical Application: These observations would provide details on micronized starch and its potential applications. Moreover, we believe that when the structures of starches were known, it is probable that the effect of VSM on the structural and physicochemical properties change of other starches might be predicted by adjusting the processing time. [ABSTRACT FROM AUTHOR]

Plants possess endless structural and chemical diversity, which is peerless with any synthetic library of small biomolecules, inspiring novel drug discovery. Plants are widely applied to encounter global health challenges such as antimicrobial resistance and diabetes. The objective of this work was to evaluate the antibacterial and antidiabetic potency of native plants grown in Cyprus. All plants were sequentially extracted with solvents of increasing polarity, namely hexane, acetone, methanol, and water. First, the phenolic and flavonoid contents of the extracts were assessed. Afterwards, the bacteriostatic and bactericidal potency of plant extracts were tested against a panel of six bacteria using the broth microdilution method, whereas the inhibitory effects on alpha-glucosidase and alpha-amylase enzymes were also determined with the employment of microplate assays. The results highlighted the superiority of Sarcopoterium spinosum as a potential enzyme inhibitor, while a knowledge base was also acquired for the inhibitory potential of all plants. Daucus carota, Ferula communis, and Tordylium.aegyptiacum displayed additionally outstanding bacteriostatic and bactericidal effects on Gram-positive bacteria at concentrations of 250 µg mL−1 and 500 µg mL−1. Overall, the present study describes the antibacterial and inhibitory activity against carbohydrate digestive enzymes of native plants grown in Cyprus delivering the first reports for many plant species. [ABSTRACT FROM AUTHOR]

The biological conversion of agro-waste biomass into value-added metabolites is one of the trendy biotechnological research areas in recent times. One of the major drawbacks of the bioprocess is the saccharification potential of the amylolytic enzyme that releases reducing sugar from complex biomass to serve as substrate for fermentation. The present study reports the production of a novel tripartite raw starch-digesting amylase (RSDA) by an indigenous Priestia flexa strain with α-, β-, and gluco-amylolytic activities and its potential for bioethanol production. Response surface statistics was employed to develop a suitable medium for improved production of the tripartite enzyme by submerged fermentation. The bioprocess selected raw starch (4.36%) Ca2+(2.71 g/L) and Zn2+ (0.0177 g/L) as significant variables which demonstrated a total RSDA activity of 7208.23 U/mL in a 5-L batch bioreactor. SDS/Native-PAGE determined the molecular weights of the 27-fold purified product as 25.2 kDa, 57.3 kDa, and 90.1 kDa for α-, β-, and gluco-amylases, respectively. Optimum temperature and pH for enzyme activity were respectively broad at 30–70 °C and 4–11. The enzyme mixture demonstrated digestibility above 90% against a variety of raw starches and simultaneous fermentation of digestate with Saccharomyces cerevisiae generated 71.69 g/L of bioethanol within 24 h suggesting great potential for bioethanologenesis. [ABSTRACT FROM AUTHOR]

Enzymatic reactions can be modulated by the incorporation of organic solvents, leading to alterations in enzyme stability, activity, and reaction rates. These solvents create a favorable microenvironment that enables hydrophobic reactions, facilities enzyme-substrate complex formation, and reduces undesirable water-dependent side reactions. However, it is crucial to understand the impact of organic solvents on enzymatic activity, as they can also induce enzyme inactivation. In this study, the enzymatic performance of Aspergillus oryzae α-amylase (Taka-amylase) in various organic solvents both experimentally and computationally was investigated. The results demonstrated that ethanol and ether sustain Taka-amylase activity up to 20% to 25% of the organic solvents, with ether providing twice the stability of ethanol. Molecular dynamics simulations further revealed that Taka-amylase has a more stable structure in ether and ethanol relative to other organic solvents. In addition, the analysis showed that the loop located near the active site in the AB-domain is a vulnerable site for enzyme destabilization when exposed to organic solvents. The ability of Taka-amylase to preserve the secondary loop structure in ether and ethanol contributed to the enzyme's activity. In addition, the solvent accessibility surface area of Taka-amylase is distributed throughout all enzyme structures, thereby contributing to the instability of Taka-amylase in the presence of most organic solvents. [ABSTRACT FROM AUTHOR]

Previous studies have shown that 2-arylbenzimidazole derivatives have a strong anti-diabetic effect. To further explore this potential, we develop new analogues of the compound using ligand-based drug design and tested their inhibitory and binding properties through QSAR analyses, molecular docking, dynamic simulations and pharmacokinetic studies. By using quantitative structure activity relationship and ligand-based modification, a highly precise predictive model and design of potent compounds was developed from the derivatives of 2-arylbenzimidazoles. Molecular docking and simulation studies were then conducted to identify the optimal binding poses and pharmacokinetic profiles of the newly generated therapeutic drugs. DFT was employed to optimize the chemical structures of 2-arylbenzimidazole derivatives using B3LYP/6-31G* as the basis set. The model with the highest R2trng set, R2adj, Q2cv, and R2test sets (0.926, 0.912, 0.903, and 0.709 respectively) was chosen to predict the inhibitory activities of the derivatives. Five analogues designed using ligand-based strategy had higher activity than the hit molecule. Additionally, the designed molecules had more favorable MolDock scores than the hit molecule and acarbose and simulation studies confirm on their stability and binding affinities towards the protein. The ADME and druglikeness properties of the analogues indicated that they are safe to consume orally and have a high potential for total clearance. The results of this study showed that the suggested analogues could act as α-amylase inhibitors, which could be used as a basis for the creation of new drugs to treat type 2 diabetes mellitus. [ABSTRACT FROM AUTHOR]

Diabetes is the third leading cause of premature death worldwide due to hyperglycemia and hyperglycemic-induced oxidative stress. The use of lactic acid bacteria (LAB) is one of the new therapeutic strategies to improve diabetes and its complication. Thus, this paper aimed to investigate the anti-hyperglycemic and antioxidant properties of the cell-free extract (CFE) and cell-free supernatant (CFS) obtained from 10 native LAB strains isolated from traditional fermented dairy products. The anti-hyperglycemic activity of LAB strains was evaluated by the α-amylase and α-glucosidase inhibition assay. The antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid, hydroxyl, superoxide anion, ferric-reducing, iron chelating, and total antioxidant capacity. Finally, phenolic and flavonoid content of CFS of samples were assessed. The findings showed that Lactobacillusdelbrueckii had the higher antidiabetic properties compared to other strains with the highest α-glucosidase inhibitory activity in CFE (25%) and CFS (50%). Other LAB strains showed high antioxidant ability in CFE and CFS. Among them, Lactobacillusparacasei had the highest antioxidant activity. The results showed that the CFS contained phenolic and flavonoid compounds. Finally, the data demonstrated that L.delbrueckii, L.paracasei, Lactobacillushelveticus, and Lactobacillusfermentum could be applied in the production of health-related products, food additives and animal feed. [ABSTRACT FROM AUTHOR]

The area of combining an enzyme and transition metals to catalyse the sequence of reactions in one-pot which improve total conversion and operational stability has seen tremendous growth in recent years. Herein, we describe the synthesis and substrate controlled modification of β-aminocarbonyl using α-amylase enzyme and Pd-based catalyst in the one-pot. Moreover, the chemo-enzymatic approach provides substituted indole derivatives unprecedentedly when isocyanide was used in the reaction. In the second phase, isocyanide was replaced by phenyl boronic acid which in turn provides the substituted amino biaryls in good yield. Next, the feasibility of one-pot chemo-enzymatic approach was proved by employing substituted 2-bromo aniline along with different isocyanide/phenyl boronic acid and obtained the corresponding products in 56–84% isolated yield. [ABSTRACT FROM AUTHOR]

In this study, the α-glucosidase (maltase-glucoamylase: MGAM) and α-amylase inhibitory properties elicited by xylooligosaccharides (XOSs) prepared from dulse xylan were analysed as a potential mechanism to control postprandial hyperglycaemia for type-2 diabetes prevention and treatment. Xylan was purified from red alga dulse powder and used for enzymatic hydrolysis using Sucrase X to produce XOSs. Fractionation of XOSs produced xylobiose (X2), β-(1→3)-xylosyl xylobiose (DX3), xylotriose (X3), β-(1→3)-xylosyl-xylotriose (DX4), and a dulse XOS mixture with n ≥ 4 xylose units (DXM). The different fractions exhibited moderate MGAM (IC50 = 11.41–23.44 mg/mL) and α-amylase (IC50 = 18.07–53.04 mg/mL) inhibitory activity, which was lower than that of acarbose. Kinetics studies revealed that XOSs bound to the active site of carbohydrate digestive enzymes, limiting access to the substrate by competitive inhibition. A molecular docking analysis of XOSs with MGAM and α-amylase clearly showed moderate strength of interactions, both hydrogen bonds and non-bonded contacts, at the active site of the enzymes. Overall, XOSs from dulse could prevent postprandial hyperglycaemia as functional food by a usual and continuous consumption. [ABSTRACT FROM AUTHOR]

Diabetes management involves preventing its risk factors. Inhibition of glucosidases and adsorption of excess free glucose are approaches to the prevention of postprandial hyperglycemia. The objective of the present study was to evaluate the antioxidant activity, glucose adsorption capacity, and α-amylase inhibitory activity in vitro of the aqueous extract of Chamaecrista nigricans. Determination of phenolic compounds content was performed using the Folin-Ciocalteu reagent and the aluminum chloride method was used for total flavonoids one. The glucose oxidase peroxidase kit was used to determine the adsorption capacity of glucose while the 3,5-dinitrosalicylic acid method was used to assess the inhibitory activity against α-amylase. Levels ranging from 33.87 ± 2.48 mg GAE/100 mg dry extract (DE) for total phenolic compounds and 1.98 ± 0.51 mg QE/100 mg DE for total flavonoids were observed. The adsorption capacity was correlated with the glucose concentration of the solution (r = 0.95) and was up to 36.61 μmol/g DE for a glucose concentration of 30 mM. The extract from the November collection was most active against α-amylase with IC50 = 0.17 mg DE/mL. Observations confirm the traditional use of this species as a preventive measure in recipes for the treatment of diabetes. This data provides a basis for future pharmaceutical prospecting. [ABSTRACT FROM AUTHOR]

Artemisia dracunculus: L. (A. dracunculus) is a popular vegetable and spice cultivated across many Middle Eastern countries. The herb’s aqueous extract has significant folkloric medicinal importance for treating various disorders. Hence, the present investigation aimed to investigate A. dracunculus hydrophilic extract phytochemical constituents and pleiotropic biological potentials, as no previous studies have investigated the antilipase and anti-αamylase effects of the A. dracunculus plant. Total phenol content and phytochemical screening assays were performed utilizing standard analytical methods. While the α-amylase inhibition, free radical-scavenging, antilipase, and cytotoxic activities were determined using dinitrosalicylic acid (DNSA), DPPH, p-nitrophenyl butyrate (PNPB), and MTS assays, respectively. The standard phytochemical analysis of A. dracunculus aqueous extract shows that this extract contains only a phenolic group. The total phenol content was 0.146 ± 0.012 mg GAE/g of the plant dry extract. The A. dracunculus aqueous extract exhibited potent DPPH free radical inhibitory (IC50 dose of 10.71 ± 0.01 μg/mL) and anti-lipase activities (IC50 dose of 60.25 ± 0.33 μg/mL) compared with Trolox (IC50 = 5.7 ± 0.92 μg/mL) and Orlistat (IC50 = 12.3 ± 0.35 μg/mL), respectively. However, it showed a weak anti-α-amylase effect (IC50 value > 1,000 μg/mL) compared with Acarbose (IC50 = 28.18 ± 1.27 μg/mL). A. dracunculus has a cytotoxic effect against the HeLa cancer cell line compared with the chemotherapeutic agent Doxorubicin. The extract has the same percent of inhibition as Doxorubicin (99.9%) at 10 mg/mL. Overall, these results pointed out for the first time the importance of considering A. dracunculus effects as a favorite candidate for preventing and treating metabolic disorders. Also, our results confirm the findings of previous reports on the role of A. dracunculus in the management of cancer and disorders resulting from the accumulation of harmful free radicals. On the contrary, the current study concluded that the antidiabetic role of A. dracunculus could be minimal. Further in-depth investigations are urgently warranted to explore the importance of A. dracunculus in pharmaceutical production. [ABSTRACT FROM AUTHOR]

The potential of native as well as acid/alkali pre-treated paddy straw was investigated as inducer substrate for Aspergillus terreus mediated cellulase and xylanase production. The cellulose, hemicellulose and lignin contents of native straw were found to be 25.94%, 17.64% and 15.82%, respectively. However, in the acid/alkali treated straw, cellulose content substantially increased to 48.09%, whereas hemicellulose and lignin contents were reported to be particularly low (11.72%, 5.08%, respectively). The FTIR spectra of native and pre-treated paddy straw further supported the variation in their chemical constituents. A. terreus repertoires were cultivated on native and treated paddy straw under solid state fermentation (SSF) and liquid fermentation (LF) conditions. The LF cultures showed higher cellulase and xylanase activities (Filter Paper cellulase: 45.33 U/L, Carboxymethyl cellulase: 104.75 U/L, β-glucosidase 80.44 U/L, Avicelase: 22.72 U/L and Xylanase: 623.69 U/L) than the respective SSF cultures. Significantly high activities of exoglucanases and endoglucanases were obtained when native straw was used as the growth substrate. The scanning electron micrographs of native and treated straw exhibited disintegration after being subjected to biological treatments, confirming the hyphal adhered growth and production of hydrolysing secretomes by the fungus. Cultivation of A. terreus on native paddy straw in LF culture system resulted in better enzyme production, and holds promise for further optimization. [ABSTRACT FROM AUTHOR]

Existing laboratory protocols for investigating the effects of factors affecting enzyme activity often require extensive hands-on manipulations and time. This can result in students either not getting the desired results or being distracted from thinking about the scientific ideas underlying the experimental designs and procedures of these protocols. In this paper, we present a lesson plan that includes a simple microscale laboratory protocol that allows students to study the action of amylase on starch and to investigate the effects of various factors (i.e., temperature, pH, substrate concentration, enzyme concentration, and competitive reversible inhibitors) on enzyme activity using immobilized amylase beads. We also show how to engage students in thinking about procedural knowledge, such as repeating measurements, measurement range, and interval. These concepts are critical to designing valid and reliable scientific investigations. [ABSTRACT FROM AUTHOR]

The crystal of 4 Bromo − 4' Chloro Benzylidene Aniline-BCBA is synthesized and crystallized in the orthorhombic with Pccn space group. The lattice constants are a = 24.995 Å, b = 6.286 Å, c = 7.326 Å. The bond parameters are reported with statistics for the comparison purpose. The computational structural analyses about Vander Waal's, 3D representation are portrayed; Raman spectroscopy provides the functional groups of BCBA. Mass spectrometer data reveal the molecular mass of BCBA as 295.5797; the Vander Waal's type of interactions presented in the title compound, a systematic study of Super-cell, Laue's model and Clinographic view of BCBA and device displays are carried out computationally. Hirshfeld surface analysis is used to envisage the many intermolecular interactions for the configuration of supramolecular structural design of BCBA. Dielectric properties are studied in the ambience of the room temperature for identifying the existence of all polarizations and in bio-medicinal use too as a better agent against diabetic macro level as IC50 of 48.76 and 52.12 for alpha-amylase and alpha-glucosidase as in-vitro case; the wear and tear of BCBA-macro is better scaled up for tribological study with frictional coefficient as 3*10−3. [ABSTRACT FROM AUTHOR]

Enzymes are biological molecules that act as catalysts and speed up the biochemical reactions. The world's biotechnological ventures are development of enzyme productiveness, and advancement of novel techniques for thriving their shelf existence. Nowadays, the most burning questions in enzyme technology are how to improve the enzyme productivity and reuse them. The immobilization of enzymes provides an excellent scope to reuse the enzymes several times to increase productivity. The main aim of the present study is the establishment of an immobilized multi-enzyme bio-system engineering process for the production of High-fructose corn syrup (HFCS) with an industrial focus. In this study, multi-enzyme such as α-amylase, glucoamylase and glucose isomerase were immobilized in various support matrices like sodium alginate, sawdust, sugarcane bagasse, rice bran and combination of alginate with cellulosic materials. The activities of the immobilized multi-enzyme system for the production of HFCS from the starch solution were determined. The multi-enzyme immobilized in sodium alginate shows better fructose conversion than free enzyme. Among the support matrices, multi-enzyme immobilized in sawdust produced total 80.74 mg/mL of fructose from starch solution and it was able to be used in several production cycles. On the other hand, multi-enzyme immobilized in combination of sodium alginate and sawdust produced the maximum amount of fructose (total 84.82 mg/mL). The free enzyme produced 17.25 mg/mL of fructose from the starch solution in only a single cycle. In this study a new fixed bed immobilized multi-enzyme bioreactor system was developed for the production of HFCS directly from starch. This finding will create a new opportunity for the application of immobilized multi-enzyme systems in many sectors of industrial biotechnology. [ABSTRACT FROM AUTHOR]

BACKGROUND: Baijiu brewing adopts the solid‐state fermentation method, using starchy raw materials, Jiuqu as saccharifying fermenting agent, and distilled spirits made by digestion, saccharification, fermentation and distillation. In the late stages of solid‐state fermentation of Baijiu, the reduced activity of glucoamylase leads to higher residual starch content in the Jiupei, which affects the liquor yield. The direct addition of exogenous glucoamylase leads to problems such as the temperature of the fermentation environment rising too quickly, seriously affecting the growth of microorganisms. RESULTS: To solve the problem of reduced activity of glucoamylase in the late stage of solid‐state fermentation of Baijiu, microcapsule beads (M‐B) based on microcapsule emulsion were prepared and the effect of M‐B on solid‐state fermentation of Baijiu was investigated. The results showed that the release of M‐B before and after drying was 53.27% and 25.77% in the liquid state (120 h) and 29.84% and 22.62% in the solid state (15 days), respectively. Adding M‐B improved the alcohol by 0.33 %vol and reducing sugar content by 0.51%, reduced the residual starch content by 1.21% of the Jiupei, and had an insignificant effect on the moisture and acidity of the Jiupei. CONCLUSION: M‐B have excellent sustained‐release properties. The addition of M‐B in solid‐state fermentation significantly increased the alcohol content, reduced the residual starch content of Jiupei, ultimately improving the starch utilization rate and liquor yield of Baijiu brewing. The preparation of M‐B provides methods and approaches for applying other active substances and microorganisms in the brewing of Baijiu. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Acidovorax avenae subsp. avenae (Aaa) is the causal agent of red stripe in sugarcane, a disease characterized by two forms: leaf stripe and top rot. Despite the importance of this disease, little is known about Aaa infection mechanisms, and studies of virulence factors (VFs) have not been reported. Previous analysis of Aaa isolates obtained from sugarcane fields in Tucumán, Argentina, suggested that pathogenicity was not related to genetic groups determined by rep‐PCR analysis. Therefore, the aims of this work were to analyse VFs produced by Aaa strains from different genetic groups and to evaluate the association among VFs, between VFs and Aaa pathogenicity and between VFs and Aaa genetic diversity. Bacterial motility (sliding, swimming and twitching), extracellular enzyme activities (proteases, amylases and endoglucanases), H2O2‐mediated oxidative stress, cell adhesion and exopolysaccharide (EPS) production were assayed in 14 Aaa isolates from different genetic groups. Pearson's correlation analysis and principal component analysis (PCA) were performed. For each VF analysed, values were variable presenting quantitative traits among the strains. Significant correlations were observed among the VFs, though no clear trend was identified between them and the pathogenicity or genetic group. Results reinforce the idea that a single VF does not account for the infection outcome, but a more complex process occurs where the interplay among VFs must be considered. The results of this study are crucial because they provide insight into the infection mechanism of this bacterium. [ABSTRACT FROM AUTHOR]

α-Amylases are essential biocatalysts representing a billion-dollar market with significant long-term global demand. They have varied applications ranging from detergent, textile, and food sectors such as bakery to, more recently, biofuel industries. Microbial α-amylases have distinct advantages over their plant and animal counterparts owing to generally good activities and better stability at temperature and pH extremes. With the scope of applications expanding, the need for new and improved α-amylases is ever-growing. However, scaling up microbial α-amylase technology from the laboratory to industry for practical applications is impeded by several issues, ranging from mass transfer limitations, low enzyme yields, and energy-intensive product recovery that adds to high production costs. This review highlights the major challenges and prospects for the production of microbial α-amylases, considering the various avenues of industrial bioprocessing such as culture-independent approaches, nutrient optimization, bioreactor operations with design improvements, and product down-streaming approaches towards developing efficient α-amylases with high activity and recyclability. Since the sequence and structure of the enzyme play a crucial role in modulating its functional properties, we have also tried to analyze the structural composition of microbial α-amylase as a guide to its thermodynamic properties to identify the areas that can be targeted for enhancing the catalytic activity and thermostability of the enzyme through varied immobilization or selective enzyme engineering approaches. Also, the utilization of inexpensive and renewable substrates for enzyme production to isolate α-amylases with non-conventional applications has been briefly discussed. [ABSTRACT FROM AUTHOR]

Salt stress is detrimental to the survival of microorganisms, and only a few bacterial species produce hydrolytic enzymes. In this study, we investigated the expression of salt stress-related genes in the salt-tolerant bacterial strain Bacillus subtilis ACP81, isolated from bamboo shoot processing waste, at the transcription level. The results indicate that the strain could grow in 20% NaCl, and the sub-lethal concentration was 6% NaCl. Less neutral protease and higher cellulase and β-amylase activities were observed for B. subtilis ACP81 under sub-lethal concentrations than under the control concentration (0% NaCl). Transcriptome analysis showed that the strain adapted to high-salt conditions by upregulating the expression of genes involved in cellular processes (membrane synthesis) and defense systems (flagellar assembly, compatible solute transport, glucose metabolism, and the phosphotransferase system). Interestingly, genes encoding cellulase and β-amylase-related (malL, celB, and celC) were significantly upregulated and were involved in starch and sucrose metabolic pathways, and the accumulated glucose was effective in mitigating salt stress. RT-qPCR was performed to confirm the sequencing data. This study emphasizes that, under salt stress conditions, ACP81 exhibits enhanced cellulase and β-amylase activities, providing an important germplasm resource for saline soil reclamation and enzyme development. [ABSTRACT FROM AUTHOR]

Amylolytic bacteria play an important role in the ecosystem, especially as probiotic and bioremediation agents in cultivation, as examples can be found in minapadi culture. The purpose of this study was to determine the amylolytic bacteria in the waters of the minapadi pond. Bacterial isolation began with bacterial sampling, inoculation and isolation of bacteria, calculation of the total abundance of bacteria, observation of bacterial morphology and bacterial purification, isolation of amylolytic bacteria, and biochemical testing. Three isolates of amylolytic bacteria were successfully obtained, namely BA5, BA6, and BA7 with a weak to moderate activity index category. The amylolytic bacterial isolates obtained were successfully grouped into 3 different groups, namely Proteus sp., Staphylococcus sp., and Micrococcus sp. Amylolytic bacteria have a biodegradation role in the minapadi cultivation environment. [ABSTRACT FROM AUTHOR]

Marennine is a blue-green pigment produced by diatoms of the Haslea genus, one of which is Haslea ostrearia. This marennine pigment is water-soluble and confirmed to contain polyphenols and glycosides. There are two forms of marennine pigment: the intracellular form of marennine (IMn) and the extracellular form of marennine (EMn). Marennine pigments exhibit various biological activities such as antibacterial, antiviral, antioxidant, allelopathic, and inhibiting the growth of other diatoms. With this biological activity, marennine can be used in various fields, one of which is aquaculture. This research aims to determine the level of toxicity and analyze the effect of exposure to the extracellular marennine produced by H. ostrearia on the juvenile blood clam Tegillarca granosa. By using the toxicity test method, Bluewater which is a supernatant of H. ostrearia (BW) was tested on juveniles of commercially important bivalve species, the blood clams Tegillarca granosa, with three treatments, namely control (0 mg/L), treatment A (0.25 mg/L) and treatment B (0.5 mg/L of BW). The observation parameters of this study were LC50-72 h with one-way ANOVA analysis. The results showed that the BW produced by H. ostrearia increased the survival of juvenile blood clams T. granosa up to 27.7% in treatment B (0.5 mg/L) compared to control within 72 h. The one-way ANOVA analysis revealed that the control and treatment B (0.5 mg/L) were significantly different. This study shows that there is potential to develop the use of marennine in shellfish aquaculture. [ABSTRACT FROM AUTHOR]

Isoxazolidine derivatives were designed, synthesized, and characterized using different spectroscopic techniques and elemental analysis and then evaluated for their ability to inhibit both α-amylase and α-glucosidase enzymes to treat diabetes. All synthesized derivatives demonstrated a varying range of activity, with IC50 values ranging from 53.03 ± 0.106 to 232.8 ± 0.517 μM (α-amylase) and from 94.33 ± 0.282 to 258.7 ± 0.521 μM (α-glucosidase), revealing their high potency compared to the reference drug, acarbose (IC50 = 296.6 ± 0.825 µM and 780.4 ± 0.346 µM), respectively. Specifically, in vitro results revealed that compound 5d achieved the most inhibitory activity with IC50 values of 5.59-fold and 8.27-fold, respectively, toward both enzymes, followed by 5b. Kinetic studies revealed that compound 5d inhibits both enzymes in a competitive mode. Based on the structure–activity relationship (SAR) study, it was concluded that various substitution patterns of the substituent(s) influenced the inhibitory activities of both enzymes. The server pkCSM was used to predict the pharmacokinetics and drug-likeness properties for 5d, which afforded good oral bioavailability. Additionally, compound 5d was subjected to molecular docking to gain insights into its binding mode interactions with the target enzymes. Moreover, via molecular dynamics (MD) simulation analysis, it maintained stability throughout 100 ns. This suggests that 5d possesses the potential to simultaneously target both enzymes effectively, making it advantageous for the development of antidiabetic medications. [ABSTRACT FROM AUTHOR]

Polyphenols were extracted from Thuja occidentalis L. bark and characterized in terms of phytochemical composition and biological activity. The optimization process investigated the impacts of extraction duration, ultrasound amplitude, and ethanol concentration on the total phenolic content in the extract. The antioxidant capacity was examined using DPPH and ABTS assays, and anti-enzyme activity against alphaglucosidase, alpha-amylase, lipase, acetylcholinesterase, and tyrosinase was determined. The experimental model revealed optimal extraction parameters: a hydroethanolic solvent with 44% v:v ethanol, a 15-minute extraction time, and a 40% ultrasound amplitude. These parameters were validated and confirmed by reaching the target Total Phenolic Content (TPC) predicted by MODDE software. The resulting lyophilized extract exhibited a high polyphenolic content (161.84 ± 23.48 mg GAE/g dry extract) and demonstrated strong antioxidant properties. Notably, it showed significant inhibition of alpha-glucosidase and alpha-amylase, suggesting potential antidiabetic effects, though its inhibition of tyrosinase activity was relatively weak. These findings provide a foundation for further investigations to confirm the qualitative and quantitative presence of specific polyphenolic metabolites in the extract and elucidate the mechanisms underlying its bioactive properties. [ABSTRACT FROM AUTHOR]

In search of 1,2,4-triazole-based antidiabetic agents, new mixed-ligand chelates of the type, [M(L)(H2O)3] (where M = Co(II), Ni(II), Cu(II) and Zn(II)), were synthesized using 2-(((3-mercapto-5-(pyridin-3-yl)-4H-1,2,4-triazol-4-yl)imino)methyl)-4-nitrophenol (H2L1) ligand and the corresponding metal acetates in 1:1 molar ratio. FTIR, NMR, mass, UV–Vis, elemental analyses, conductivity measurements, ESR, XRD, TGA, SEM, and EDAX studies were used to determine the nature of bonding, coordination characteristics and the stability of compounds. Spectral and physico-analytical studies suggest that the ligand functioned in a binegative tridentate manner with phenolic-O, azomethine-N and thiol-S as the coordinating sites and an octahedral geometry was proposed for the metal complexes. The quantum computational calculations of the synthesized compounds were executed in the ground state using DFT/B3LYP level with 6-311++G as basis set. In vitro alpha-amylase and alpha-glucosidase inhibitory assay revealed moderate to good results for the synthesized compounds and especially the complex 3 against alpha-amylase and complex 4 against alpha-glucosidase were found to be effective inhibitors. Additionally, the compounds were explored for molecular docking studies against the human pancreatic alpha-amylase (PDB id: 1BSI) and alpha-glucosidase (PDB id: 5ZCC). The molecular docking calculations of complex 3 into 1BSI revealed a binding energy of − 8.6 kcal/mol, whereas that of complex 4 against 5ZCC resulted in a binding energy of − 7.4 kcal/mol respectively, both of which exceeds that of the standard Acarbose. New Schiff base ligand and its transition metal chelates were synthesized. Furthermore, the synthesized compounds were examined for in vitro alpha-amylase and alpha-glucosidase inhibitory activity. [ABSTRACT FROM AUTHOR]

Using non‐conventional starch can benefit the industry since it can present different properties. It also can lead to new properties upon physical modification, which improves its derivate film properties. Therefore, the aim of this work is to evaluate the heat–moisture treatment (HMT) on pinhão starch and its effectiveness in film properties. After isolation using water as a solvent, the pinhão starch is treated by HMT for 16 h at 110 °C. Native and HMTed starches are used to produce biodegradable films. Pinhão starch and starch films chemical and physical properties are properly characterized. The HMT causes some changes in short‐range ordered structures, reduces the relative crystallinity, and shifts the pinhão starch from C‐type to A‐type. Also, HMT decreases the peak viscosity and the breakdown, and improves thermal stability. These starch changes upon HMT reduces water vapor permeability, increases tensile strength, and elongation at the break of pinhão starch films. Desirable changes in starch and film properties are achieved by physically modifying pinhão starch using HMT, which is a promising alternative to chemical modifications. [ABSTRACT FROM AUTHOR]

α-Amylase inhibitors constitute a category of compounds capable of diminishing α-amylase activity and hold significant promise for the development of novel drugs. This study focused on assessing the potential of Schisandra chinensis Baill extract to inhibit α-amylase. However, sieving and segregating inhibitors from intricate mixtures presents an obstacle. Consequently, this study devised and refined a methodology centered on immobilized α-amylase to identify and isolate natural inhibitors from S. chinensis. The immobilized α-amylase, featuring a load capacity of 254.18 ± 1.20 μg/mg and a specific activity of 2.69 ± 0.01 U/mg, was achieved through meticulous optimization of preparation conditions. Subsequently, this immobilized enzyme was used to isolate bioactive compounds from S. chinensis extract. The results revealed the presence of an active compound with an IC 50 of 324.12 ± 11.23 μg/mL. This compound was isolated via magnetic immobilized enzyme-based ligand fishing, employing high-speed countercurrent chromatography, and was conclusively identified as 5-hydroxymethylfurfural. The molecular docking analyses revealed its ability to interact with the active pocket of α-amylase through multiple hydrogen bonds. [Display omitted] • α-Amylase was immobilized on mesoporous silica particles for ligand fishing. • Main active component was screened from S. chinensis by immobilized α-amylase. • 5-Hydroxymenthyfurfural was the main active component in S. chinensis fruit. • The interaction between active compound and α-amylase was investigated. [ABSTRACT FROM AUTHOR]

Starch hydrolyzing amylases are among the vital enzymes used in the food, textile, detergent, and pharmaceutical industries. The aim was to isolate amylase bacteria from agricultural soil, its extraction and subsequent purification. Within the scope of this work, using a 1% starch agar medium, potent thermostable bacterial amylase (A-4) was isolated from agricultural soil samples. Molecular analysis revealed the amylase-producing bacterial strain (A-4) to be Citrobacter portucalensis. We used agro-industrial-based wastes (wheat bran, potato peel, and sugarcane bagasse) in amylase production. Culture condition optimization showed that high amylase activity of 4.96 U/mL was obtained at temperature 55 ◦C and pH 5.0 after 72 h of incubation. The bestsuited substrate was nitrogen, and carbon sources were wheat bran, yeast, and lactose for enzyme production. In addition, metal ions Ca+2 and Mg+2 showed a net positive effect on the rate of the enzymatic reaction, and Al+3 and Co+2 inhibited the activity. Protein purification results demonstrated a 4-fold increase with 13% yield and specific activity of 40 U/mL/min. SDS-PAGE indicated the molecular weight of partially purified protein as ~56 kDa. The stain removal efficiency of the purified enzyme showed considerable results. The finding revealed the use of low-cost and readily available substrates, making C. portucalensis (A-4) a promising candidate for detergent and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Current work reports the highly selective, solvent‐free synthesis of an endorsed bioactive compound, Bis benzylidine cyclohexanone (BBC) through solid acid catalysed cross aldol condensation route and checks its in‐vitro bio activity. The catalytic support (Multiwalled carbon nanotube) employed was synthesized through the highly sophisticated catalytic chemical vapor deposition (CVD) method and simple mechanical grinding strategy was adopted to decorate sulfonic acid on the support. The C1s X‐ray photoelectron peak of at 290.3 eV confirms the effective interaction of sulfonic acid with MWCNT. The sharp and intense desorption peaks observed at approximately 528.7 °C and 655.15 °C in the TPD analysis unmistakably substantiate the strong acidity of the synthesized system. The alpha amylase inhibition activity of the synthesized compound was calculated to be around 88.5 %, which is in par with the commercial drug as it could inhibit only 96 %. Further, the in‐silico (Docking and Molecular Dynamic Simulation) investigations unveiled a new target site for the compound and this can further be studied in detail to advance the applications in drug design. Detailed scrutiny of various parameters was conducted to validate the bioactivity and pharmaceutical potential of the synthesized compound. [ABSTRACT FROM AUTHOR]

The current study aims to evaluate and characterize the probiotic andantidiabetic properties of lactic acid bacteria (LAB) obtained from milk and other dairy-based products. The strains were tested physiologically, biochemically, and molecularly. Based on biochemical tests and 16S rRNA gene amplification and sequencing, all three isolates RAMULAB18, RAMULAB19, and RAMULAB53 were identified as Lacticaseibacillus paracasei with homology similarity of more than 98%. The inhibitory potential of each isolate against carbohydrate hydrolysis enzymes (a-amylase and a-glucosidase) was assessed using three different preparations of RAMULAB (RL) isolates: the supernatant (RL-CS), intact cells (RL-IC), and cellfree extraction (RL-CE). Additionally, the isolate was evaluated for its antioxidant activity against free radicals (DPPH and ABTS). The strain's RL-CS, RL-CE, and RL-IC inhibited a-amylase (17.25 to 55.42%), a-glucosidase (15.08-59.55%), DPPH (56.42-87.45%), and ABTS (46.35-78.45%) enzymes differently. With the highest survival rate (>98%) toward tolerance to gastrointestinal conditions, hydrophobicity (>42.18%), aggregation (>74.21%), as well as attachment to an individual's colorectal cancer cell line (HT-29) (>64.98%), human buccal and chicken crop epithelial cells, all three isolates exhibited extensive results. All three isolates exhibited high resistance toward antibiotics (methicillin, kanamycin, cefixime, and vancomycin), and other assays such as antibacterial, DNase, hemolytic, and gelatinase were performed for safety assessment. Results suggest that the LAB described are valuable candidates for their significant health benefits and that they can also be utilized as a beginning or bio-preservative tradition in the food, agriculture, and pharmaceutical sectors. The LAB isolates are excellent in vitro probiotic applicants and yet additional in vivo testing is required. [ABSTRACT FROM AUTHOR]

Inhibiting α-amylase can lower postprandial blood glucose levels and delay glucose absorption, offering an effective approach for the development of antidiabetic diets. In this study, an active constituent with inhibitory activity against α-amylase was isolated and purified by bioassay-guided fractionation from Carya cathayensis Sarg. peel (CCSP). The active constituent was identified by NMR and Q-Exactive Orbitrap Mass Spectrometry as 5-O-p-coumaroylquinic acid (5-CQA). 5-CQA possessed strong inhibitory activity against α-amylase, with an IC50 value of 69.39 µM. In addition, the results of the kinetic study indicated that 5-CQA was a potent, reversible, noncompetitive inhibitor against α-amylase. The findings indicate that 5-CQA derived from CCSP has potential as a novel inhibitor against α-amylase, which can help mitigate postprandial blood sugar spikes, making it suitable for inclusion in antidiabetic diets. [ABSTRACT FROM AUTHOR]

Chilo partellus is a serious pest of agricultural crops. The present investigation was carried out to characterize the gut α-amylase from C. partellus and to explore its inhibition kinetics with various α-amylase inhibitors. α-amylase eluted at ve/vo of 1.38–1.48 on sephadex G-100 column chromatography with 15.58 folds purification. It had a single isoform of 65 KDa with an optimum pH of 8.0. It was thermally stable up to 50 °C and showed the highest activity with starch. A critical analysis of Lineweaver–Burk plot showed that the Michaelis constant, Km, of α-amylase for starch was 0.32 mg/ml while Vmax was observed to be 1.47 nmoles of reducing sugars formed/min/ml of enzyme. Among the various chemicals studied, citric acid, salicylic acid, oxalic acid, zinc chloride and calcium nitrate were found to be the potent inhibitors of α-amylase activity. Through a concerted study of the primary and secondary plots of Lineweaver–Burk, fractional velocity and combination plots, it was revealed that citric acid, salicylic acid, oxalic acid, zinc chloride and calcium nitrate caused complete inhibition of α-amylase activity by mixed- non-competitive-uncompetitive mode. [ABSTRACT FROM AUTHOR]

This study aims to evaluate the potential of filamentous fungi consortiums to design a bioprocess for producing hydrolytic enzymatic cocktails on lignocellulosic biomass. Four different microorganisms were analyzed in solid and submerged monoculture fermentation systems: Aspergillus niger, Trichoderma harzianum, Rhizopus oryzae, and Thermomyces lanuginosus. A. niger and R. oryzae were selected as the most suitable to form a consortium. Two different inoculation strategies were assessed, one was simultaneous and the other was a 12-h post-phase inoculation. Xylanase, cellulase, amylase, pectinase, and protease activities were evaluated in mono and mixed cultures. Furthermore, the secretomes of the fermentation systems were analyzed by mass spectrometry to compare and determine the difference in the protein profiles excreted. The mixed culture of A. niger and R. oryzae, which were inoculated simultaneously in solid-state fermentation, showed the highest levels of enzyme at 96 h, 30°C, 1 × 107 conidia, and 65% relative humidity. The proportion of xylanase (98 IU/gds), cellulase (27 IU/gds), amylase (30 IU/gds), pectinase (21 IU/gds) and protease (108,000 IU/gds) was enhanced by 25% regarding the monoculture, demonstrating that the use of microbial consortiums can be a promising alternative to obtain enzymatic cocktails with high synergism. [ABSTRACT FROM AUTHOR]

In the present research, the impact of fermentation with two strains of Lactiplantibacillus plantarum subsp. plantarum (PTCC 1896 and PTCC 1745) on physicochemical properties, antioxidant bioactive compounds, and some health‐promoting features of guava juice was investigated. Results showed a significant (p <.05) decrease in pH, total soluble solids, glucose and fructose residues, vitamin C, and total carotenoids after 32 h of fermentation. Total phenolic content, free radical scavenging abilities, and ferrous reducing power were markedly enhanced during the fermentation process. Moreover, fermented juice represented good enzyme inhibition abilities (α‐amylase and α‐glucosidase) and anti‐inflammatory activities. The initial amount of angiotensin‐converting enzyme inhibitory activity (26.5%) increased to 72.1% and 66.4% in L. plantarum subsp. plantarum 1896 and L. plantarum subsp. plantarum 1745 treatments, respectively. These findings reveal that guava juice fermentation with the studied Lactobacillus strains can be a promising strategy to augment the functional properties of the fruit‐based beverage. [ABSTRACT FROM AUTHOR]

α-Amylase is a generally acknowledged molecular target of a distinct class of antidiabetic drugs named α-glucosidase inhibitors. This class of medications is scarce and rather underutilized, and treatment with current commercial drugs is accompanied by unpleasant adverse effects. However, mammalian α-amylase inhibitors are abundant in nature and form an extensive pool of high-affinity ligands that are available for drug discovery. Individual compounds and natural extracts and preparations are promising therapeutic agents for conditions associated with impaired starch metabolism, e.g., diabetes mellitus, obesity, and other metabolic disorders. This review focuses on the structural diversity and action mechanisms of active natural products with inhibitory activity toward mammalian α-amylases, and emphasizes proteinaceous inhibitors as more effective compounds with significant potential for clinical use. [ABSTRACT FROM AUTHOR]

The insect pathogenic fungus, Ascosphaera apis, is the causative agent of honeybee chalk brood disease. Amylases are secreted by many plant pathogenic fungi to access host nutrients through the metabolism of starch, and the identification of new amylases can have important biotechnological applications. Production of amylase by A. apis in submerged culture was optimized using the response surface method (RSM). Media composition was modeled using Box–Behnken design (BBD) at three levels of three variables, and the model was experimentally validated to predict amylase activity (R2 = 0.9528). Amylase activity was highest (45.28 ± 1.16 U/mL, mean ± SE) in media composed of 46 g/L maltose and1.51 g/L CaCl2 at a pH of 6.6, where total activity was ~11-fold greater as compared to standard basal media. The enzyme was purified to homogeneity with a 2.5% yield and 14-fold purification. The purified enzyme had a molecular weight of 75 kDa and was thermostable and active in a broad pH range (> 80% activity at a pH range of 7–10), with optimal activity at 55 °C and pH = 7.5. Kinetic analyses revealed a Km of 6.22 mmol/L and a Vmax of 4.21 μmol/mL·min using soluble starch as the substrate. Activity was significantly stimulated by Fe2+ and completely inhibited by Cu2+, Mn2+, and Ba2+ (10 mM). Ethanol and chloroform (10% v/v) also caused significant levels of inhibition. The purified amylase essentially exhibited activity only on hydrolyzed soluble starch, producing mainly glucose and maltose, indicating that it is an endo-amylase (α-amylase). Amylase activity peaked at 99.38 U/mL fermented in a 3.7 L-bioreactor (2.15-fold greater than what was observed in flask cultures). These data provide a strategy for optimizing the production of enzymes from fungi and provide insight into the α-amylase of A. apis. [ABSTRACT FROM AUTHOR]

Bacillus amyloliquefaciens α‐amylase (BAA) is one of well‐known midrange thermostability amylases that are widely used in food and washing processes. However, the improvement of its catalytic properties by molecular modification is still lagging. To improve the activity of α‐amylase BAA, mutants BAA28 and BAA294 are constructed via error‐prone PCR and purified by column chromatography in the present study. The catalytic efficiencies (Kcat/Km) of BAA28 and BAA294 are 2.42 and 2.73 mL mg−1 s−1, which are 43% and 61% higher than that of the wild‐type BAA, respectively. Their specific activities are also increased by 40% and 62%, respectively, with no apparent changes of optimum temperature and pH. Homology modeling and molecular docking analysis suggest that the reduced steric hindrance is an important factor that enhances catalytic efficiencies and specific activities of the variants. These results may deepen the understanding of the mechanisms underlying the effects of each mutation on the catalytic efficiency of BAA and facilitate the construction of potent BAA mutants. [ABSTRACT FROM AUTHOR]

In this study, we conducted research on the preparation of aerogels using cellulose and starch as the primary materials, with the addition of N,N′-methylenebisacrylamide (MBA) as a cross-linking agent. The chemical, morphological and textural characteristics of the aerogels were found to be influenced by the proportions of cellulose, starch, and cross-linking agent that were utilized. An increase in the proportion of cellulose led to stronger adsorption forces within the aerogel structure. The aerogel showed a fine mesh internal structure, but the pores gradually increased with the further increase in cellulose. Notably, when the mass fractions of starch and cellulose were 5 wt% and 1 wt% respectively, the aerogels exhibited the smallest pore size and largest porosity. With an increase in the crosslinking agent, the internal structure of the aerogel first became dense and then loose, and the best internal structure was displayed at the addition of 3 wt%. Through texture analysis and the swelling test, the impact of the proportion of cellulose and MBA on the aerogel structure was significant. Dye adsorption experiments indicated that MBA affected the water absorption and expansion characteristics of the aerogel by improving the pore structure. Lastly, in tests involving the loading of vitamin E, the aerogels exhibited a higher capacity for incorporating vitamin E compared to native starch. [ABSTRACT FROM AUTHOR]

In this study, some new sulfonylurea derivatives based on the sulfanilamide compound were synthesized. IR, 1H-NMR, 13C-NMR spectroscopic methods and elemental analysis data were used to confirm the structures of the synthesized compounds. ABTS, DPPH, Cuprac and β-Carotene/linoleic acid assays were performed to evaluate the antioxidant activities of sulphonylurea derivatives. The activities of these compounds against some enzymes (acetylcholinesterase, butrylcholinesterase, tyrosinase, α-amylase and α-glucosidase) were also investigated. Compounds S7, S8, S13 and S14 showed inhibitory activity against α-amylase enzyme with IC50 values of 227.84±1.48- 298.27±8.61 µM. In addition, the drug-likeness properties and solubility of the compounds were determined by computational programs. [ABSTRACT FROM AUTHOR]

A novel spirooxindole‐pyrrolidine clubbed thiochromene and pyrazole motifs were synthesized by [3+2] cycloaddition (32CA) reactions in one step process starting from the ethylene‐based thiochromene and pyrazole scaffolds with the secondary amino‐acids and substituted isatins in high yield. The 32CA reaction of AY 10 with ethylene derivative 6 has also been studied with Molecular Electron Density Theory. The high nucleophilic character of AY 10, N = 4.39 eV, allows explaining that the most favorable TS‐on is 13.9 kcal mol−1 below the separated reagent. This 32CA, which takes place through a non‐concerted one‐step mechanism, presents a total ortho regio‐ and endo stereoselectivity, which is controlled by the formation of two intramolecular H...O hydrogen bonds. The design of spirooxindole‐pyrrolidines engrafted thiochromene and pyrazole was tested for alpha‐amylase inhibition and show a high efficacy in nanoscale range of reactivity. The key interaction between the most active hybrids and the receptor was studied by molecular docking. The physiochemical properties of the designed spirooxindole‐pyrrolidines were carried out by in silico ADMET prediction. The newly synthesized most potent hybrid could be considered as a lead compound for drug discovery development for type 2 diabetes mellitus (T2DM). [ABSTRACT FROM AUTHOR]